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利用抑制性消减杂交技术对埃及伊蚊精巢中的减数分裂驱动表型进行转录谱分析。

Transcript profiling of the meiotic drive phenotype in testis of Aedes aegypti using suppressive subtractive hybridization.

机构信息

Eck Institute for Global Health, Department of Biological Sciences, University of Notre Dame, Notre Dame, IN 46556-5645, USA.

出版信息

J Insect Physiol. 2011 Sep;57(9):1220-6. doi: 10.1016/j.jinsphys.2011.05.014. Epub 2011 Jun 14.

Abstract

The meiotic drive gene in Aedes aegypti is tightly linked with the sex determination locus on chromosome 1, and causes highly male-biased sex ratios. We prepared cDNA libraries from testes from the Ae. aegypti T37 strain (driving) and RED strain (non-driving), and used suppressive subtraction hybridization techniques to enrich for T37 testes-specific transcripts. Expressed sequence tags (ESTs) were obtained from a total of 2784 randomly selected clones from the subtracted T37 (subT37) library as well as the primary libraries for each strain (pT37 and pRED). Sequence analysis identified a total of 171 unique genes in the subT37 library and 299 unique genes among the three libraries. The majority of genes enriched in the subT37 library were associated with signal transduction, development, reproduction, metabolic process and cell cycle functions. Further, as observed with meiotic drive systems in Drosophila and mouse, a number of these genes were associated with signaling cascades that involve the Ras superfamily of regulatory small GTPases. Differential expression of several of these genes was verified in Ae. aegypti pupal testes using qRT-PCR. This study increases our understanding of testes gene expression enriched in adult males from the meiotic drive strain as well as insights into the basic testes transcriptome in Ae. aegypti.

摘要

埃及伊蚊的减数分裂驱动基因与染色体 1 上的性别决定基因紧密连锁,并导致高度偏向雄性的性别比例。我们从埃及伊蚊 T37 品系(驱动)和 RED 品系(非驱动)的睾丸中制备 cDNA 文库,并使用抑制性消减杂交技术富集 T37 睾丸特异性转录本。从 T37 消减文库(subT37)和每个品系的初级文库(pT37 和 pRED)中总共随机选择了 2784 个克隆,获得了表达序列标签(EST)。序列分析鉴定了 subT37 文库中总共 171 个独特基因和三个文库中 299 个独特基因。在 subT37 文库中富集的大多数基因与信号转导、发育、生殖、代谢过程和细胞周期功能有关。此外,与果蝇和小鼠的减数分裂驱动系统一样,这些基因中的许多与涉及 Ras 调节小 GTP 酶超家族的信号级联有关。使用 qRT-PCR 在埃及伊蚊蛹睾丸中验证了其中几个基因的差异表达。这项研究增加了我们对减数分裂驱动品系中雄性成虫睾丸中富集的基因表达的理解,并深入了解埃及伊蚊的基本睾丸转录组。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c139/3167017/2089d0b28b9b/nihms-305413-f0001.jpg

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